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Biomedical subjects

H Kitagawa

Publications and source records attributed to H Kitagawa.

At least 127 records · Page 7Linked to original sources
PubMed1999

Adenovirus-mediated gene transfer of glial cell line-derived neurotrophic factor prevents ischemic brain injury after transient middle cerebral artery occlusion in rats.

To examine a possible protective effect of exogenous glial cell line-derived neurotrophic factor (GDNF) gene expression against ischemic brain injury, a replication-defective adenoviral vector containing GDNF gene (Ad-GDNF) was directly injected into the cerebral cortex at 1 day before 90 minutes of transient middle cerebral artery occlusion (MCAO) in rats. 2,3,5-Triphenyltetrazolium chloride staining showed that infarct volume of the Ad-GDNF-injected group at 24 hours after the transient MCAO was significantly smaller than that of vehicle- or Ad-LacZ-treated group. Enzyme-linked immunosorbent assay (ELISA) for immunoreactive GDNF demonstrated that GDNF gene products in the Ad-GDNF-injected group were higher than those of vehicle-treated group at 24 hours after transient MCAO. Immunoreactive GDNF staining was obviously detected in the cortex around the needle track just before or 24 hours after MCAO in the Ad-GDNF group, whereas no or slight GDNF staining was detected in the vehicle group. The numbers of TUNEL, immunoreactive caspase-3, and cytochrome c-positive neurons induced in the ipsilateral cerebral cortex at 24 hours after transient MCAO were markedly reduced by the Ad-GDNF group. These results suggest that the successful exogenous GDNF gene transfer ameliorates ischemic brain injury after transient MCAO in association with the reduction of apoptotic signals.

Adenoviruses, Human↗
PubMed1999

Expression of nerve growth factor in hepatolithiasis.

AIMS/BACKGROUND: Nerve growth factor (NGF) has recently been shown to influence the survival and function of non-neuronal inflammatory cells, possibly through its activity as a colony-stimulating factor. It may also play an important role in acute inflammation and tissue repair. However, no prior report has focused on NGF in chronic inflammatory diseases of the gastrointestinal and biliary tracts. The aim of this study was to examine the expression of NGF in hepatolithiasis. METHODS: Twenty-six liver specimens resected from 22 patients with intrahepatic calcium bilirubinate stones and from 4 patients with intrahepatic cholesterol stones were examined immunohistochemically. RESULTS: The 22 patients with calcium bilirubinate stones demonstrated NGF immunoreactivity associated with surrounding inflammatory cells that was localized to the epithelia of proliferative peribiliary glands in the ductal wall. However, neither the surface lining of the bile duct nor hepatocytes expressed detectable NGF immunoreactivity. In the cholesterol stones cases in contrast, peribiliary glandular elements and inflammatory cell infiltration were less extensive than those observed in cases of calcium bilirubinate stones, and NGF immunoreactivity was not noted. CONCLUSIONS: These observations suggest that proliferative peribiliary glands express NGF protein in chronic proliferative cholangitis. This is characteristic of intrahepatic calcium bilirubinate stones.

Adult↗
PubMed1999

Apoptosis of villous epithelial cells and follicle-associated epithelial cells in chicken cecum.

The process of the disappearance of epithelial cells was examined in chicken cecal villi and follicle-associated epithelium (FAE). The apoptotic epithelial cells with intense DNA-fragmentation and their exfoliation were found in the villous tips. The epithelial cells with weak DNA-fragmentation were seen in the upper portion of the villi and their sparse exfoliations were also found there. Numerous epithelial cells in the intestinal lumen expressed the apoptotic features. A row of apoptotic epithelial cells with DNA-fragmentation was also found in the apical FAE, whereas no M cells exhibited any apoptotic signs. In all cecal regions, CD3+, CD8+, and TCR2+ lymphocytes were predominant in the epithelium at the upper portion of the villi and the FAE. CD4+ lymphocytes were mainly seen in the lamina propria. TCR1+ lymphocytes were not abundant in comparison with TCR2+ lymphocytes in the epithelium. TCR3+ T lymphocytes were rarely detected. These results suggest that the chicken cecal epithelial cells exfoliated into the lumen after the induction of the apoptosis, and that the induction may be involved with CD3+, CD8+, and TCR2+ lymphocytes. No death in M cells suggests that M cells may transform into microvillous epithelial cells.

Animals↗
PubMed1999

Method of lymphocytotoxic crossmatch test for feline renal transplantation.

The optimal condition for methods of lymphocytotoxic crossmatch test for feline renal transplantation was investigated. On separation of viable lymphocytes from whole blood, the best results were obtained when Ficoll-diatrizoate with 1.078 of a specific gravity at 20 degrees C was centrifuged with 800 x g for 30 min at 4 degrees C. A nylon wool column was used to separate T and B cells from lymphocyte fraction. The ratio of T cells in nylon wool effluent cells was 95%, while the ratio of B cells in adherent cells was 41%. Lymphocytotoxic crossmatch tests were performed by using the effluent cells as T cells and the adherent cells as B cells, at 37 degrees C (warm) and 4 degrees C (cold). The ratio of B cells in adherent cells was low, however, the result was utilized as a matching test before transplantation by combining with the T cell result. The trypan blue stain method made it easier than the eosin stain method to distinguish living and dead cells. The lymphocytotoxic crossmatch tests were performed on 15 pairs of healthy cats, and only one pair showed doubtful positive against anti-B cell cold antibodies. During acute rejection after renal transplantation in two pairs which were negative on any anti-lymphocyte antibodies before the transplantation, the anti-T cell warm antibodies became positive in both pairs, and the anti-T cell cold antibodies became positive on one of the two pairs.

Animals↗
PubMed1999

Quantitative studies of the optic nerve fiber layer in the chicken retina.

The optic nerve fiber layer (NFL) of the chicken retina was studied quantitatively and morphologically at 17 positions along seven radially arranged bands from the dorsal tip of the pecten oculi using electron microscopy. The number of nerve fibers was counted in areas 6 microm in width x the full thickness of the NFL. Myelinated nerve fibers in the NFL were also identified immunohistochemically using anti-myelin basic protein serum. The dorsal area (dorsal, dorso-temporal and dorso-nasal bands) in the retina had thin NFL and contained the largest number of nerve fibers, which were mainly thin and unmyelinated. The ventral area (ventral and ventro-temporal bands) had a thick NFL and contained a relatively small number of nerve fibers, many of which were myelinated. The nasal band had the thickest NFL and contained as many nerve fibers as the dorsal area, with the temporal band showing a high ratio of myelinated fibers. The band had a thick NFL and contained many nerve fibers with a relatively low ratio of myelinated fibers. The relationship between the number and composition of nerve fibers in the NFL to the chicken visual characteristics was discussed. Although the myelin in the chicken retina was loose type, the myelin-forming cells were similar in appearance to dense oligodendrocytes. retina, morphometry, myelinated fiber, nerve fiber layer.

Animals↗
PubMed1999

Concentrations of serum granulocyte-colony-stimulating factor in normal pregnancy and preeclampsia.

OBJECTIVE: In order to elucidate the potential role of granulocyte-colony-stimulating factor (G-CSF) during the course of normal pregnancy and preeclampsia, we measured the serum concentrations of G-CSF in both normal pregnant women and preeclamptic patients. METHODS: Sera of 10 nonpregnant women, 34 normal pregnant women (n = 10, first trimester; n = 10, second trimester; n = 14, third trimester), 10 postpartum women, 10 mild preeclamptic patients, and 10 severe preeclamptic patients were collected. The serum concentrations of G-CSF were measured by enzyme immunoassay. RESULTS: The serum level of G-CSF in normal pregnant women (third trimester: 38.3 +/- 15.3 pg/mL; mean +/- SD) was significantly increased when compared with the levels observed in nonpregnant women (20.3 +/- 10.1 pg/mL, p < 0.05), which was similar to the G-CSF concentrations in postpartum subjects (20.7 +/- 9.1 pg/mL). The mild and severe preeclamptic patients showed significantly higher levels of G-CSF (56.9 +/- 18.8 pg/mL, p < 0.05; 73.2 +/- 30.4 pg/mL, p < 0.0001, respectively) than those noted in the third trimester women. The preeclamptic patients who presented with edema had greater (p < 0.01) serum levels of G-CSF (75.5 +/- 25.0 pg/mL) compared with nonedematous patients (44.7 +/- 14.9 pg/mL). The serum levels of G-CSF significantly correlated with both weight gain (p < 0.05), diastolic blood pressure (p < 0.01), and systolic blood pressure (p < 0.01), but not with white blood cell counts. CONCLUSION: Serum concentrations of G-CSF are increased in normal and, even more so, in preeclamptic pregnancies. Because there was no relationship between serum G-CSF concentration and the number of leukocytes, G-CSF might act not to promote the physiological leukocytosis of pregnancy, but to stimulate the function of leukocytes such as phagocytosis. Moreover, it might be that G-CSF plays important roles in the activation of granulocytes or vascular endothelial injury, which are considered to be important pathological conditions in the development of preeclampsia.

Adult↗
PubMed1999

Activated lansoprazole inhibits cancer cell adhesion to extracellular matrix components.

Integrins play an important role in tumor dissemination. The purpose of this study was to examine whether a SH-reactive reagent inhibits the adhesion of the human pancreatic cancer cell line AsPC-1 to extracellular matrix components. Activated lansoprazole (AG-2000) was used as the SH-reactive reagent because this compound is known to react with SH groups but does not permeate the cell membrane. The effect of AG-2000 on the adhesion of AsPC-1 cells to matrix was examined, using both an in vitro adhesion assay and an in vivo nude mouse xenograft model of peritoneal implantation. In the in vitro adhesion assay, a 60-min exposure of AsPC-1 cells to AG-2000 resulted in a dose-dependent inhibition of AsPC-1 cell adhesion to laminin, fibronectin and type IV collagen, although AG-2000 did not affect the viability of AsPC-1 cells by MTT assay. In the in vivo assessment of AsPC-1 cell implantation, the AsPC-1 cells were initially preincubated with AG-2000 for 60 min to ensure adequate exposure of the AsPC-1 cells to AG-2000 before intraperitoneal injection. AG-2000 significantly inhibited the peritoneal implantation of the AsPC-1 cells in nude mice. These findings suggest that a short exposure of cancer cells to AG-2000 can inhibit cancer cell adhesion to extracellular matrix components.

Animals↗
PubMed1999

Frigen II improves the reliability of measurement of interleukin-1 related substances in amniotic fluid.

BACKGROUND: To investigate the role of interleukin-1 related substances in amniotic fluid in normal term labor without intrauterine infection. METHODS: Amniotic fluid samples were collected from forty-one patients with various backgrounds. A novel pre-assay treatment using Frigen II was introduced to improve the recovery rates of cytokines, i.e., interleukin-1alpha, interleukin-1beta and interleukin-1 receptor antagonist, prior to ELISA assay. Urine samples from newborn infants were also tested. RESULTS: The concentrations of interleukin-1alpha, interleukin-1beta and interleukin-1ra were significantly higher in samples from normal vaginal delivery. The higher levels of interleukin-1alpha were also observed in samples from preterm labor without infection. Preterm infants produced more of interleukin-1 receptor antagonist in urine compared with term infants. CONCLUSIONS: This study provides evidence of the possible involvement of interleukin-1 related substances in labor without signs of infection. The data from newborn urine suggests that amniotic interleukin-1ra originates from the fetus. However, amniotic fluid interleukin-1alpha and interleukin-1beta may be derived from maternal tissue, such as decidua.

Amniotic Fluid↗
PubMed1999

[Anesthetic management of patients with malignant pleural effusion undergoing hyperthermic perfusion under thoracoscopy].

Nine patients with malignant pleural effusion due to lung cancer had been scheduled for hyperthermic treatment with warmed distilled water (40 degrees C) under thoracoscopy. This treatment aims to produce adhesion of the lungs to reduce pleural effusion. To evaluate the risk of general anesthesia for patients with lung cancer at the end stage, we examined the problems of perioperative management. Seven out of nine patients were classified into ASA physical status > or = III and seven patients into Hugh Jones > or = III Shapiro's score was > or = 5 in four patients. The average %VC was 60 +/- 16 and % FEV1.0 was 41 +/- 18% (means +/- SE). A double lumen endotracheal tube was inserted and anesthesia was maintained with inhalational anesthetics. In two cases, one-lung ventilation could not be maintained because of severe hypoxemia during hyperthermic perfusion. Hypertension occurred in three cases and hypotension in one by direct heat stimulation of the cardiopulmonary system. Although their preoperative risk was poor, there were no major complications and the quality of life was improved. We stress that careful anesthetic management is important for avoiding hypoxemia and hemodynamic instability during this treatment.

Aged↗
PubMed1999

[Surgery for metastatic brain and spinal tumor from colorectal cancer].

UNLABELLED: The results of invasive treatment of brain metastasis of colorectal cancer (6 cases) and spinal metastasis of colorectal cancer (3 cases), aimed at alleviating the symptoms, were analyzed. In all 6 cases of brain metastasis, metastases were also found in some other remote sites (often the liver). The average period from resection of the primary lesion to resection of the brain metastasis was 3 years and 3 months for this group. Of these 6 cases, 5 had cerebral metastasis and 1 had cerebellar metastasis. The preoperative performance status (PS) was 1 in 1 case and 2 in 5 cases. After surgery, the PS improved in all 6 cases, allowing all the patients to be discharged. There was only one patient whose death was attributable to brain metastasis. In all 3 cases of spinal metastasis, metastases were also detected in the lungs or liver (synchronous in) 1 case, metachronous in 2 cases). The preoperative PS was 2 in 1 case, 3 in 1 case and 4 in 1 case. After surgery, the PS improved in 2 cases. One of the three patients is still alive. CONCLUSION: The results of this study suggest that it is not impossible to improve the QOL of patients with brain or spinal metastasis of cancer by surgical intervention, although the response may vary from patient to patient.

Brain Neoplasms↗
PubMed1999

Biochemical and physiological properties of pedicellarial lectins from the toxopneustid sea urchins.

Pedicellarial lectins (SUL-I, SUL-II, and TGL-I) were purified from the toxopneustid sea urchins, Toxopneustes pileolus and Tripneustes gratilla using gel filtration chromatography, affinity chromatography, and reverse-phase HPLC. SUL-I (Nakagawa et al., 1996) and SUL-II from the large globiferous pedicellariae of T. pileolus are D-galactose-binding lectins with molecular masses of 32 kDa and 23 kDa, respectively; while TGL-I from the globiferous pedicellariae of T. gratilla is a Ca(2+)-independent heparin-binding lectin with a molecular mass of 23 kDa. SUL-I induced mitogenic stimulation on murine splenocytes but TGL-I did not. At higher dose ranges SUL-I exhibited inhibitory effects on the cells. The dual response to SUL-I was effectively inhibited by D-galactose. SUL-I enhanced norepinephrine-induced contraction of isolated rat mesenteric artery with endothelium. When endothelium was removed from the artery, acetylcholine did not relax the norepinephrine-induced contraction. In the same artery the enhancing effect of the contraction by SUL-I was abolished, suggesting that SUL-I acts on the endothelium of mesenteric artery, and may release prostanoids. The present results suggest an extracellular function for SUL-I that may have wide-ranging effects in physiological processes. The primary role of pedicellarial lectins from T. pileolus and T. gratilla might be defense against a foreign body.

Agglutination↗
PubMed1999

Oral administration of leflunomide (HWA486) results in prominent suppression of immunoglobulin E formation in a rat type 1 allergy model.

Leflunomide, a drug being developed for use in the treatment of rheumatoid arthritis, was evaluated in an ovalbumin (OVA)-induced rat type 1 allergy model. In a dose of 1 mg/kg/day, it strongly suppressed the formation of OVA-specific IgE, thus preventing the elevation of the plasma histamine level and induction of anaphylactic shock observed after i.v. challenge with OVA. Studies on leflunomide's effects on the secondary antibody response showed that administration during the primary immune response remarkably diminished the secondary antibody responses, except IgM, even without further drug administration. Furthermore, when administered during the secondary response after rechallenge, both the total IgE and OVA-specific IgE serum levels declined rapidly to nearly baseline levels. Although OVA-specific IgG1, IgG2a, and IgM did not decrease from their primary response levels, these classes' secondary responses were strongly suppressed. In an in vitro study, the proliferation and antibody production of OVA-stimulated spleen cells, derived from Brown Norway rats, were strongly inhibited by A77 1726, leflunomide's active metabolite. When uridine was added to the cell culture, this molecule's effect on cell proliferation was completely restored, whereas the antibody production was partially restored. These findings are consistent with data indicating that leflunomide is a dihydroorotate dehydrogenase inhibitor. Taken together, the above findings suggest the therapeutic potential of leflunomide against type 1 allergic diseases.

Administration, Oral↗
PubMed1998

Functional expression and genomic structure of human chondroitin 6-sulfotransferase.

The cDNA and gene encoding human chondroitin 6-sulfotransferase (C6ST) have been cloned. The expression of a soluble recombinant form of the protein in COS-1 cells produced an active sulfotransferase, which used as acceptor substrates polymer chondroitin, various chondroitin sulfate isoforms and chondroitin sulfate tetrasaccharides. The identification of the reaction products demonstrated that the enzyme transferred sulfate to position 6 of GalNAc in the GlcAbeta1-3GalNAc but not the IdoAalpha1-3GalNAc nor the GlcAbeta1-3GalNAc(4-O-sulfate) sequences. The human C6ST gene spans more than 20 kb and consists of three exons. The protein-coding domain of the C6ST gene is divided into two discrete exons.

Amino Acid Sequence↗
PubMed1998

Demonstration of the immature glycosaminoglycan tetrasaccharide sequence GlcAbeta1-3Galbeta1-3Galbeta1-4Xyl on recombinant soluble human alpha-thrombomodulin. An oligosaccharide structure on a "part-time" proteoglycan.

Thrombomodulin (TM), a cell surface glycoprotein, is a critical mediator of endothelial anticoagulant defenses occurring both as a chondroitin sulfate proteoglycan (beta-TM) and a protein (alpha-TM) unsubstituted by chondroitin sulfate (CS), hence its description as a "part-time" proteoglycan (PG) (Fransson, L. A. (1987) Trends Biochem. Sci. 12, 406-411). Sugar analysis was performed on alpha-TM to investigate a possible biosynthetic mechanism for part-time PGs. Recombinant human alpha-TM, which was expressed in CHO-K1 cells, separated by anion-exchange chromatography from beta-TM, and purified by immunoaffinity chromatography (Nawa, K., Sakano, K., Fujiwara, H., Sato, Y., Sugiyama, N., Teruuchi, T., Iwamoto, M., and Marumoto, Y. (1990) Biochem. Biophys. Res. Commun. 171, 729-737), was used for analysis. Preliminary sugar composition analysis after acid hydrolysis showed Xyl in addition to Gal, GalNAc, GlcNAc, Man, Fuc, and Glc. O-Glycosidically-linked oligosaccharides were liberated by mild alkaline treatment and purified. The isolated oligosaccharide fraction was derivatized with a fluorophore 2-aminobenzamide (2AB), resulting in two fluorescent components, a 2AB-oligosaccharide and a putative 2AB-Glc. Based on structural analysis by a combination of sequential exoglycosidase digestion and 500-MHz 1H NMR spectroscopy of the 2AB-oligosaccharide, the structure of the oligosaccharide was elucidated as GlcAbeta1-3Galbeta1-3Galbeta1-4Xyl, which turned out to represent a glycosaminoglycan (GAG)-protein linkage region tetrasaccharide common to various PGs and was considered to be a biosynthetic intermediate of an immature GAG chain. The results may indicate that at least one class of the so-called part-time PGs bear the linkage tetrasaccharide at the GAG attachment sites and that the critical determining step or the rate-limiting step for PG biosynthesis is the transfer of the fifth sugar residue, the first hexosamine, rather than xylose.

Animals↗
PubMed1998

Comparison of laboratory test results before and after surgical removal of heartworms in dogs with vena caval syndrome.

OBJECTIVE: To compare results of laboratory tests in dogs with vena caval syndrome before and after surgical removal of heartworms. DESIGN: Longitudinal uncontrolled clinical trial. ANIMALS: 51 dogs with vena caval syndrome. PROCEDURE: Heartworms were removed from the area of the tricuspid valve and pulmonary arteries via venotomy and by use of flexible alligator forceps. Blood samples were obtained before and 10 days after removal of heartworms. Red and white blood cell counts were determined, using an automated cell counter. Biochemical tests were performed, using a dry chemical method. RESULTS: 45 dogs survived the procedure, and 6 died or were euthanatized after surgical treatment. After surgery, RBC count and total protein, albumin, calcium, and sodium concentrations increased, and total bilirubin, ammonia, BUN, creatinine, uric acid, and potassium concentrations decreased in dogs that survived. Creatine kinase, L-lactate dehydrogenase, and amylase activities decreased, whereas alkaline phosphatase and gamma-glutamyltransferase activities and total cholesterol concentration increased. Alanine transaminase activity decreased in 27 dogs but increased in 3 dogs. Changes in test results in dogs that did not survive. Significant differences were found in RBC count, gamma-glutamyltransferase activity, and total protein, total cholesterol, BUN, and total bilirubin concentrations before and after removal of heartworms. CLINICAL IMPLICATIONS: Hepatic and renal functions improve rapidly after surgical removal of heartworms, presumably because general and pulmonary circulation is normalized. However, cholestasis may develop, and dogs that survive may need additional treatment to preserve hepatic function.

Anemia↗
PubMed1998

Induction of DNA fragmentation and HSP72 immunoreactivity by adenovirus-mediated gene transfer in normal gerbil hippocampus and ventricle.

Foreign genes have been successfully transferred and expressed in experimental animal brains using adenoviral vectors. However, it is not fully understood whether adenovirus-mediated gene transfer causes stressful or cytotoxic injury in brain. A replication-defective adenoviral vector containing the Escherichia coli lacZ gene (AdCMVnLacZ) was directly injected into right hippocampus and lateral ventricle of normal gerbil brains. Temporal and spatial profiles of the expression of lacZ gene products, DNA fragmentation detected by terminal deoxynucleotidyl d-UTP nick end labeling (TUNEL) staining, and heat shock protein 72 (HSP72) immunoreactivity were examined until 21 days after the injection. In the ventricle, lacZ gene was immediately and strongly expressed at 8 hr after the injection of AdCMVnLacZ, with a peak at 1-3 days, and disappeared by 21 days. Although a small number of choroid plexus cells were TUNEL positive at 3 and 7 days, no HSP72 immunostaining was observed in the ventricle. Small-to-moderate expression of lacZ gene was found in the needle route from 8 hr to 3 days after the injection, and a small number of TUNEL-positive cells were detected at the needle track at 1-3 days. In the hippocampus, lacZ gene was markedly expressed around the dentate gyrus (DG) at 8 hr to 3 days with a peak at 1 day. Large number of TUNEL or moderate-to-dense HSP70 staining cells were also detected in the same area. CA1 neuronal cells just adjacent to the needle route showed TUNEL positivity at 1 to 3 days. However, the TUNEL staining was not associated with lacZ gene expression. The majority of lacZ-expressing cells were discriminated from the TUNEL-positive cells, whereas some were double-positive with HSP72 staining in DG. Cellular loss was observed in the CA1 layer around the needle route. An apoptotic change was morphologically observed in the marginal region of the DG at 1-3 days and in the ventricle at 3-7 days. In the sham control group, TUNEL-positive or HSP72-staining cells were only detected around the needle track including CA1 cells adjacent to the needle route. These data suggest that adenoviral gene transfer may induce direct traumatic injury in the CA1 sector near the needle route, indirect apoptotic cell loss in the DG and ventricle, and stressful effect on the dentate granule cells in association with adenovirus infection in normal gerbil brain.

Adenoviridae↗